TPR-MET oncogenic rearrangement: detection by polymerase chain reaction amplification of the transcript and expression in human tumor cell lines.
Activation of the MET protooncogene by a rearrangement involving the fusion of TPR and MET specific gene sequences has been observed in a human osteosarcoma cell line (HOS) treated in vitro with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). No information has been available about the possible occurrence of this rearrangement in human tumors. To facilitate rapid screening of human cell lines and tumor samples for this specific gene rearrangement, we developed a sensitive detection method based on polymerase chain reaction (PCR) amplification of TPR-MET mRNA. cDNA was generated from cellular transcripts by using one of the PCR primers, which was then used as a template for PCR amplification of a 205-base-pair region carrying the breakpoint. An end-labeled internal probe was hybridized in solution to an aliquot of the PCR product for detecting amplification. Cells could be directly screened by the assay without prior isolation of RNA. A 205-base-pair DNA fragment characteristic of the TPR-MET rearrangement was detected in cell lines previously known to contain this altered sequence. The rearrangement was also detected at very low levels in the parental (nontransformed) cell line, HOS TE-85. A preliminary survey of cell lines derived from a variety of human tumors indicates that TPR-MET rearrangement occurred and was expressed at very low frequencies by cells from 7 of 14 tumors of nonhematopoietic origin.
Soman
N R
NR
Division of Toxicology, Whitaker College of Health Sciences and Technology, Cambridge, MA.
Wogan
G N
GN
Rhim
J S
JS
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002878:Chromosomes, Human, Pair 1; D002897:Chromosomes, Human, Pair 7; D004273:DNA, Neoplasm / Q000235:genetics; D015870:Gene Expression; D005786:Gene Expression Regulation; D015321:Gene Rearrangement; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000138:chemical synthesis; D012516:Osteosarcoma; D016133:Polymerase Chain Reaction; D011519:Proto-Oncogenes; D012334:RNA, Neoplasm / Q000235:genetics; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured / Q000187:drug effects
D004273:DNA, Neoplasm; D015345:Oligonucleotide Probes; D012334:RNA, Neoplasm; D008769:Methylnitronitrosoguanidine